ZFIN Image: Zhou et al., 2015, Fig. 4

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Figure Caption

Fig. 4 (A) Microarray analysis of mRNA expression in E10.5 Nfatc1^Cre;Krit1^fl/fl and Krit1^fl/+ littermate hearts reveals increased levels of Klf2, Klf4, KLF2 target genes and Adamts5, and reduced levels of Dll4 and Tmem100. N = 4 for both genotypes. (B) qPCR of E10.5 hearts reveals preserved or increased expression of Nrg1 and FGF growth factors following endocardial Krit1 loss. (C) qPCR of E10.5 hearts reveals elevated levels of Klf2 and established KLF2 target genes following endocardial Krit1 loss. (D) qPCR analysis of genes associated with cardiac jelly matrix proteins and matrix-degrading proteases in E10.5 hearts reveals elevated levels of Adamts5 following endocardial Krit1 loss. N = 3 for Krit1fl/fl, N = 4 for Nfatc1^Cre;Krit1fl/+, N = 5 for Nfatc1^Cre;Krit1fl/fl in (B)–(D). (E) In situ hybridization for Klf2 in E10.5 Nfatc1^Cre;Krit1^fl/fl and Krit1^fl/fl littermate hearts. (F) Immunostaining for KLF4 protein (arrows) and myocardium (MF20) in E10.5 Nfatc1^Cre;Krit1^fl/fl and Krit1^fl/fl littermate hearts. (G) Immunoblot analysis of KLF2 protein in whole E10.5 Nfatc1^Cre;Krit1fl/fl and Nfatc1Cre;Krit1fl/+ and Krit1fl/fl littermate hearts. GAPDH is shown as a loading control. (H) Immunostaining using anti-DPEAAE antibody to detect ADAMTS-cleaved versican reveals increased levels in the E10.5 Nfatc1^Cre;Krit1fl/fl heart. Boxed regions are shown at higher magnification on the right. (I) Immunoblot analysis of lysate derived from whole E10.5 hearts reveals increased levels of cleaved versican (DPEAAE) and the ADAMTS5 protease with endocardial loss of Krit1. GAPDH is shown as a loading control. Scale bars represent 100 μm. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

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Acknowledgments

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Reprinted from Developmental Cell, 32, Zhou, Z., Rawnsley, D.R., Goddard, L.M., Pan, W., Cao, X.J., Jakus, Z., Zheng, H., Yang, J., Arthur, J.S., Whitehead, K.J., Li, D., Zhou, B., Garcia, B.A., Zheng, X., Kahn, M.L., The cerebral cavernous malformation pathway controls cardiac development via regulation of endocardial MEKK3 signaling and KLF expression, 168-80, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell